Team:Paris-Saclay/Project/Notebook/Week 11
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- | | style="width: 50%;"| | + | | style="width: 50%;"|Determination of the concentration of plasmid pSB1A2 which has been linearized by HINDIII. We are expecting a band at size (2079+935) ~3kbp |
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- | | style="width: 50%;"|A new PCR of the plasmid pSB1A2 is realized to obtain more quantity of the plasmid. We are expecting a band at | + | | style="width: 50%;"|A new PCR of the plasmid pSB1A2 is realized to obtain more quantity of the plasmid. We are expecting a band at size 2079 bp. |
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PCR program used: | PCR program used: | ||
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====14th August==== | ====14th August==== |
Revision as of 00:37, 27 September 2012
Week 11
13th August
Purification by PCR clean-up of BBa_K115017 treated with DPNI. To visualize the fragment, an electrophoresis has been made with a 2% Agarose gel. We are expecting a band at the size of 123 bp. | |
New PCR of BBa_K115017 to obtain more quantity of the fragment |
- Digestion by DPNI of the BBa_K115017 in order to degrade the matrix plasmid.
Determination of the concentration of plasmid pSB1A2 which has been linearized by HINDIII. We are expecting a band at size (2079+935) ~3kbp | |
A new PCR of the plasmid pSB1A2 is realized to obtain more quantity of the plasmid. We are expecting a band at size 2079 bp. |
14th August
Determination of the concentration of the BBa_K274100 by electrophoresis on a gel at O.8% Agarose. We are expecting a band at 2100bp size |
- Digestion of the BBa_K274100 by DPNI to degrade the matrix plasmid.
- Stocking up cells with glycerol
- PSB1A3Amil CP + Ampicilline
- PSB1C3 Amil GFP + Chloramphinicol
15th August
Day of public holiday in France.
16th August
PCR of BBa_K274100 and visualization by 0.8% Agarose gel electrophoresis. We are expecting a band at 3400bp. |
- Digestion of the BBa_K274100 by DPNI to degrade the matrix plasmid.
17th august
Visualization of the DPNI’s digestion of the plasmid pSB1A2 by 0.8% Agarose gel electrophoresis. . We are expecting a band at the size of 2079 bp. |
- Miniprep of the plasmid pSB1A2.
- Miniprep of BBa_K274100 and BBa_K115017
- Gibson assembly of the B construction
- Transformation of DH5α competent cells with the Gibson B construction. The Petri dishes are placed at 37°C.
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